1-aryl-1-hydroxy-2,3-diamino-propyl amines, 1-heteroaryl-1-hydroxy-2,3-diamino-propyl amines and related compounds having analgesic and/or immuno stimulant activity

ABSTRACT

Compounds of the formula 
     
       
         
         
             
             
         
       
     
     where the variables have the meaning defined in the specification have analgesic and/or immunostimulant effect in mammals.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a continuation of Ser. No. 11/814,601, filed Mar.17, 2008, which is a §371 National Stage of PCT/US2006/002570, filedJan. 25, 2006, which claims the benefit of U. S. Provisional ApplicationSer. No. 60/647,271, filed Jan. 26, 2005, each of which is herebyincorporated by reference herein.

FIELD OF THE INVENTION

The present invention relates to derivatives of1-aryl-1-hydroxy-2,3-diamino-propyl amines,1-heteroaryl-1-hydroxy-2,3-diamino-propyl amines and to relatedcompounds having analgesic and in some cases immunostimulant activity.

The present invention also relates to pharmaceutical compositionscontaining these compounds as active ingredient for alleviating oreliminating pain in mammals and/or stimulating the immune system inmammals and to methods of using said pharmaceutical compositions asanalgesics and/or immunostimulants.

BACKGROUND ART

1-Phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) was discoveredby Vunam, R. R. and Radin, N., Chem. Phys. Lipids, 26, 265-278, 1980.Preparation of PDMP is described in Inokuchi, J. et al., J. Lipid Res.28, 565-571, 1987; Radin, A. et al., NeuroProtocols, 3(2), 145-55, 1993;Radin, A. et al., J. Lipid Res. 36, 611-621, 1995 and U.S. Pat. No.5,916,911.

These derivatives inhibit glucosylcer//amide (GlcCer) formation byinhibiting the enzyme GlcCer synthase, thereby lowering the level ofglycosphingolipids.) The isomers most active have theR,R-(D-threo)-configuration. Four enantiomers are produced during thesynthesis. Because only the D-threo enantiomers are active in inhibitingthe glucosylceramide synthase, resolution of the active D-threoinhibitors was performed by chiral chromatography.

Moreover, D-threo-PDMP has antitumor activity via inhibition ofglycosphingolipid biosynthesis as described by Inokuchi J., CancerLetters 38(1-2), 23-30, 1987.

Furthermore, it was also reported that D-threo-PDMP suppresses synapticfunction by Mizutani A. et al., Biochem. Biophys. Res. Commun, 222,494-498, 1996.

Preparation of enantiomerically pure D-threo-PDMP has been reported byMitchell, Scott A. [J. Org. Chem., 63 (24), 8837-8842, 1998]; Miura, T.et al, [Bioorg. Med. Chem., 6, 1481-1498, 1998]; Shin, S. et al.,[Tetrahedron asymmetry, 11, 3293-3301, 2000]; WO 2002012185.

L-threo-PDMP is an agent for treating neuronal diseases WO 95/05177.This compound is also described to be an agent for protecting brain inU.S. Pat. No. 6,407,064. Moreover treatment with L-threo-PDMP aftertransient forebrain ischemia in rats ameliorated the deficit of a welllearned spatial memory by an 8-arm maze task, suggesting a potential forneurodegenerative disorders as described by Inokuchi et al., Ann. N.Y.Acad. Sci., 845(1), 219-224, 1998 and JP 10324671 (Seikagaku Kogyo

Co.).

A stereoselective synthesis of enantiomerically pure D-threo-PDMP hasalso been described by Shin, S. et al., Tetrahedron asymmetry, 11,3293-3301, 2000 and WO 2002012185 the key step is the regioselectivecleavage by nitrogen nucleophiles, as morpholine, of the C(3)-N-bond ofnon-activated enantiomerically pure aziridine-2-methanols.

On the other hand, the synthesis of enantiomerically pure(1S,2S)-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (L-threo-PDMP)from L-serine has also been described by Mitchell, Scott A., J. Org.Chem., 63 (24), 8837-8842, 1998.

Other known methods to obtain L-threo-PDMP are described by Miura, T. etal, Bioorg. Med. Chem., 6, 1481-1498, 1998 and in JP-A-9-216858.

L-threo-PDMP is an agent for treating neuronal diseases WO 95/05177.This compound is also described to be an agent for protecting brain inU.S. Pat. No. 6,407,064. Moreover treatment with L-threo-PDMP aftertransient forebrain ischemia in rats ameliorated the deficit of a welllearned spatial memory by an 8-arm maze task, suggesting a potential forneurodegenerative disorders as described by Inokuchi et al., Ann. N.Y.Acad. Sci., 845(1), 219-224, 1998 and JP 10324671 (Seikagaku Kogyo

Co.).

Synthesis of (1S,2S)-threo- and(1R,2S)-erythro-1-phenyl-2-palmitoylamino-3-N-morpholino-1-propanol(PPMP) were described starting from Garner aldehyde of L-serine, byNishida, A., Synlett, 4, 389-390, 1998.

Compounds with longer chain fatty acyl groups (than decanoyl) have beenfound to be substancially more effective as inhibitor of GCS.D-threo-1-phenyl-2-palmitoylamino-3-pyrrolidino-1-propanol (P4 or PPPP)analogues were first obtained by a Mannich reaction as described Abe, A.et al., J. Biochem., 111, 191-196, 1992 or U.S. Pat. No. 5,916,911 andWO 2001004108.

Preparation of D-threo-4′-hydroxy-P4, one of the most potent inhibitorof GCS, was described by Lee, L. et al., J. Biol. Chem., 274, 21,14662-14669, 1999. In addition, a series of dioxane substitutions wasdesigned and tested. These included3′,4′-methylenedioxyphenyl-3′,4′-ethylenedioxyphenyl-, and3′,4′-trimethylenedioxyphenyl-substituted homologues.

Synthesis of enantiomerically pureD-threo-1-phenyl-2-benzyloxycarbonylamino-3-pyrrolidino-1-propanol(PBPP) and D-threo-P4 and its analogues fromN-benzyloxycarbonyl-D-serine, was described by Jimbo M. et al, J.Biochem., 127(3), 485-91, 2000 and EP 782992 (Seikagaku Kogyo Co.). PBPPis described as a potent GCS inhibitor.

Novel prodrugs of P4 derivatives were described in US 20020198240 and WO2002062777.

Synthesis of enantiomerically pure of D-threo-ethylenedioxy-P4 andD-threo-p-methoxy-P4 were described by Husain A. and Ganem B.,Tetrahedron Lett., 43, 8621-8623, 2002. The key step is a highlysyn-selective additions of aryl Grignard reagents to Garner aldehyde.

Diastereoselective synthesis of P4 analogues were described in U.S. Ser.No. 03/0,153,768 and WO 2003045928 (Genzyme Corp.); Oxazolines I[R1=(un)substituted aryl; R², R³═H, (un)substituted aliphatic;NR²R³=heterocyclic] are prepared as intermediates for P4glucosyltransferase inhibitors from R¹CHO and R²R³NCOCH₂CN. Thus, methylisocyanoacetate CNCH₂CO₂Me was treated with pyrrolidine and the amidewas treated with 1,4-benzodioxane-6-carboxaldehyde, followed byhydrolysis of the oxazoline using HCl in methanol, reduction of the ketogroup of amide II using LiAlH₄, and acylation with palmitoyl chloride togive D,L-threo-ethylenedioxy-P4 III.

Synthesis of enantiopure P4 analogues were described in WO 2003008399(Genzyme Corp.).

P4 derivatives, such as I [R¹, R⁵=un(substituted) aromatic; R², R³═H,un(substituted) aliphatic; NR²R³=(un)substituted non-aromaticheterocyclic ring; R⁴═O, H₂], were prepared for their therapeutic use asGCS inhibitors. Thus, D-threo-ethylenedioxy-P4 was prepared via amultistep synthetic sequence starting from S-(+)-Ph glycinol,phenyl-α-bromoacetate, 1,4-benzodioxan-6-carboxaldehyde, pyrrolidine andpalmitoyl chloride.

New D-threo-P4 analogues that bear ether substituents on the aromaticring have been recently synthesized from D-serine and found to suppressneurite extension in an embryonic insect cell line as described bySlavish., J. P. et al., Bioorg. Med. Chem. Lett., 14, 1487-1490, 2004.

Further references which serve as background to the present inventionare U.S. Pat. Nos. 5,945,442; 5,952,370; 6,030,995 and 6,051,598;Journal of Labelled Compounds & Radiopharmaceuticals (1996), 38(3),285-97; Published PCT application WO 01/38228; and Kastron et al.Latvijas PSR Zinatnu Akademijas Vestis, Kimijas Serija (1965) (4),474-7.

Significantly, according to the best knowledge of the present inventorsnone of the compounds of the prior art which are structurally similar tothe novel compounds of the present invention are known in the prior artas analgesics or immuno stimulants.

SUMMARY OF THE INVENTION

The present invention is directed to compounds of Formula 1

where R₁ is H or alkyl of 1 to 6 carbons,

-   R₂ is H, alkyl of 1 to 6 carbons or the R₁ and R₂ groups together    with the nitrogen form a saturated or unsaturated 4, 5, 6 or 7    membered ring that optionally includes one or two heteroatoms    independently selected from N, O and S, said 4, 5, 6 or 7 membered    ring optionally being substituted with a halogen, COOH, CH₂OH, OH,    B(OH)₂, cyano or with an alkyl group having 1 to 6 alkyl groups;-   R₃ is independently selected from H, alkyl of 1 to 20 carbons, aryl    or heteroaryl, aryl-alkyl or heteroaryl-alkyl where the alkyl moiety    has 1 to 4 carbons, cycloalkyl of 3 to 6 carbons, said aryl or    heteroaryl groups being optionally substituted with 1 to 3 groups    independently selected from the group consisting of halogen, alkyl    of 1 to 6 carbons, alkoxy of 1 to 6 carbons and thioxy of 1 to 6    carbons, or-   R₃ is CO—R₇ or CO—O—R₇ where R₇ is H, alkyl of 1 to 20 carbons,    benzyl, alkyl of 1 to 20 carbons substituted with and NH₂ group,    with a NHCOOalkyl or with an NH—COalkyl group where the alkyl group    has 1 to 6 carbons, or R₇ is aryl, heteroaryl, aryl-alkyl or    heteroaryl-alkyl where the alkyl moiety is branched or unbranched    and has 1 to 4 carbons, said aryl or heteroaryl groups being    optionally substituted with 1 to 3 groups independently selected    from the group consisting of halogen, alkyl of 1 to 6 carbons,    alkoxy of 1 to 6 carbons and thioxy of 1 to 6 carbons;-   R₄ is H, alkyl of 1 to 6 carbons or CO—R₈ where R₈ is alkyl of 1 to    6 carbons;-   the wavy lines represent bonds connected to carbons having R or S    configuration, and R₁₀ is selected from the groups of formulas (i)    and (ii)

where the * indicates the carbon atom to which the remaining moiety ofthe molecule is attached;

-   R₅ and R₆ independently are H, alkyl of 1 to 6 carbons, halogen,    alkoxy of 1 to 6 carbons or the R₅ and R₆ groups together with the    atoms to which they are attached jointly form a carbocyclic or a    heterocyclic ring, the carbocyclic ring having 5 or 6 atoms in the    ring, the heterocyclic ring having 5 or 6 atoms in the ring and 1 to    3 heteroatoms independently selected from N, O and S, and-   said carbocyclic or heterocyclic ring jointly formed by R₅ and R₆    being optionally substituted with 1 to 6 R₉ groups where R₉ is    independently selected from halogen, alkyl of 1 to 6 carbons, alkoxy    of 1 to 6 carbons,-   with the proviso:-   that when R₁₀ has formula (ii) then Formula 1 does not include    compounds where R₄ is hydrogen and R₁ and R₂ jointly with the    nitrogen form a morpholin or a pyrrolidin ring and where R₅ and R₆    both are H or one of R₅ and R₆ is OCH₃ and the other is H, and the    present invention is also directed to all pharmaceutically    acceptable salts of said compounds.

The present invention is also directed to pharmaceutical compositionscontaining the above-noted novel compound to be used as analgesicsand/or immuno stimulants in mammals, and to methods of using saidpharmaceutical compositions as analgesics and/or as immuno stimulants.

DETAILED DESCRIPTION OF THE INVENTION

A general description of the compounds of the invention is provided inthe Summary Section of the present application for patent. Mostcompounds of the invention contain one or more asymmetric centers, suchthat the compounds may exist in enantiomeric as well as indiastereomeric forms. In fact, most of the compounds of the presentinvention have two asymmetric carbons adjacent to one another andtherefore can exist in erythro or threo form, with each of these twoforms having dextrorotatory (D) or levorotary (L) enantiomers. Althoughthe threo form is generally preferred in accordance with the presentinvention for analgesic activity, unless it is specifically notedotherwise, the scope of the present invention includes all enantiomers,diastereomers and diastereomeric and racemic mixtures. In light of theforegoing, it should be clearly understood that the designation “DL” or“(+/−)” or “(±)” in this application includes the pure dextrorotatoryenantiomer, the pure levorotatory enantiomer and all racemic mixtures,including mixtures where the two enantiomers are present in equal or inunequal proportions. Moreover, for simplicity sake in many of thestructural formulas, such as in the example below, only one of theenantiomers is actually shown but when the designation “DL” (or “(+/−)”or “(±)”) appears it also includes the enantiomeric form (mirror image)of the structure actually shown in the formula.

For Example:

Thus, in the example above, only one enantiomer is shown, but becausethe designation “DL” (or “(+/−)” or “(±)”) appears below the formula,its optical isomer

and all racemic mixtures of the two optical isomers are also included.

In the case of some compounds of the present invention one enantiomer ofthe threo, and in some cases of the erythro, is significantly moreactive as an analgesic or immuno stimulant than the other enantiomer ofthe same pair. For this reason the isolated enantiomer which issignificantly more active than the other is considered a novel andinventive composition even if the racemic mixture or one singleenantiomer of the same compounds have already been described in theprior art.

Some of the novel compounds of the present invention may contain threeor more asymmetric centers.

Keeping the foregoing examples in mind a person of ordinary skill in theart should readily understand the scope of each described example,although in a broad sense all isomers, enantiomers and racemic mixturesare within the scope of the invention.

The term “alkyl” in the general description and definition of thecompounds includes straight chain as well as branch-chained alkylgroups.

Generally speaking the compounds of the invention may form salts withpharmaceutically acceptable acids or bases, and such pharmaceuticallyacceptable salts of the compounds of Formula 1 are also within the scopeof the invention.

Referring now to the novel compounds of Formula 1, the R₅ and R₆ groupspreferably both are independently selected from H, alkyl, alkoxy andstill more preferably are H. In the preferred compounds the R₃ groupsare preferably both H, or one of the R₃ groups is H and the other is anacyl group or an arylalkylcarbamoyl group. The R₄ group is preferably H(but see the “proviso” in the Summary section) or alkanoyl, and the R₁and R₂ groups preferably are pyrrolidino or morpholino.

The presently most preferred novel compounds of the invention aredisclosed with their structural formulas in the ensuing Table and/ordescription, showing activity of exemplary compounds relevant to theirability to act as analgesics.

Biological Activity, Modes of Administration

The novel compounds of the invention have analgesic and/orimmunostimulant activity in mammals. Some of the compounds described inthe introductory section which per se are known in the art have beendiscovered by the present inventors to also have analgesic effect inmammals. To the best of the knowledge of the present inventors theanalgesic or immunostimulant biological activity of the known compoundswas not known before the present discovery.

An art-accepted model or assay for measuring an analgesic effect of acompound in chronic pain (in particular peripheral neuropathy) is themodel known as Kim and Chung 1992, Pain 150, pp 355-363 (Chung model).This model involves the surgical ligation of the L5 (and optionally theL6) spinal nerves on one side in experimental animals. Rats recoveringfrom the surgery gain weight and display a level of general activitysimilar to that of normal rats. However, these rats developabnormalities of the foot, wherein the hindpaw is moderately everted andthe toes are held together. More importantly, the hindpaw on the sideaffected by the surgery appears to become sensitive to low-thresholdmechanical stimuli and will perceive pain instead of the faint sensationof touch. This sensitivity to normally non-painful touch, called“tactile allodynia”, develops within the first week after surgery andlasts for at least two months. The allodynia response includes liftingthe affected hindpaw to escape from the stimulus, licking the paw andholding it in the air for many seconds. None of these responses isnormally seen in the control group.

To produce the tactile allodynia, rats are anesthetized before surgery.The surgical site is shaved and prepared either with betadine orNovacaine. Incision is made from the thoracic vertebra XIII down towardthe sacrum. Muscle tissue is separated from the spinal vertebra (leftside) at the L4-S2 levels. The L6 vertebra is located and the transverseprocess is carefully removed with a small rongeur to expose the L4-L6spinal nerves. The L5 and L6 spinal nerves are isolated and tightlyligated with 6-0 silk thread. The same procedure is done on the rightside as a control, except no ligation of the spinal nerves is performed.

After a complete hemostasis is confirmed, the wounds are sutured. Asmall amount of antibiotic ointment is applied to the incised area, andthe rat is transferred to the recovery plastic cage under a regulatedheat-temperature lamp.

On the day of the experiment, at least seven days after the surgery,typically six rats per test group are administered the test drugs byintraperitoneal (i.p.) injection or oral gavage (p.o.). For i.p.administration, the compounds are formulated in H₂O and given in avolume of 1 ml/kg body weight by injecting into the intraperitonealcavity. For p.o. administration, the compounds are formulated in H₂O andgiven in a volume of 1 ml/kg body weight using an 18-gauge, 3 inchgavage needle that is slowly inserted through the esophagus into thestomach.

Tactile allodynia is assessed via von Frey hairs, which are a series offine hairs with incremental differences in stiffness. Rats are placed ina plastic cage with a wire mesh bottom and allowed to acclimate forapproximately 30 minutes. To establish the pre-drug baseline, the vonFrey hairs are applied perpendicularly through the mesh to themid-plantar region of the rats' hindpaw with sufficient force to causeslight buckling and held for 6-8 seconds. The applied force has beencalculated to range from 0.41 to 15.1 grams. If the paw is sharplywithdrawn, it is considered a positive response. A normal animal willnot respond to stimuli in this range, but a surgically ligated paw willbe withdrawn in response to a 1-2 gram hair. The 50% paw withdrawalthreshold is determined using the method of Dixon, W. J., Ann. Rev.Pharmacol. Toxicol. 20:441-462 (1980) hereby incorporated by reference.Tactile allodynia is measured prior to and 15, 30, and 60 minutes afterdrug administration. The post-drug threshold is compared to the pre-drugthreshold and the percent reversal of tactile sensitivity is calculatedbased on a normal threshold of 15.1 grams.

Table 1 below indicates the degree of pain reversal obtained in theChung model with exemplary compounds of the invention. Theintraperitonial (i.p.) and/or intravenous (iv) administration of thecompounds was in doses ranging from 1 μg/kg to 300 μg/kg or 3 mg/kg POand the peak percentage of reversal of allodynia was measured at 15, 30or 60 minutes after administration, as is indicated in the table. Dataare expressed as the highest % allodynia reversal (out of 3 time points:15 min, 30 min, or 60 min. post-drug) with a minimum of a 20% allodyniareversal in the rat Chung model. Comparisons between groups (drugtreated vs. saline treated) were made using a two-tailed, 2-sample,unpaired t-test. Compounds that are not shown which were notstatistically analgesic following an IP dose of 300 ug/kg, but may stillbe analgesic. Compounds that do not exhibit significant analgesia at 100mg/kg are not considered to be analgesic.

TABLE 1 Peak % Pain Compound reversal: time post Dost μg/kg, # ChcmicalFormula dose Mode of administ. 1

30 min  44% 300 μg/kg IP 3

60 min  92% 300 μg/kg IP 46 

60 min  96%  30 μg/kg IP

An art accepted method for measuring immuno stimulation comprisessystemic administration of compounds to assay for the ability tostimulate the immune system, possibly due to nonspecific upregulation ofthe hemolymphoreticular system. This upregulation could result inincreased numbers of lymphocytes of both T- and B-cell lineage. Althoughapplicant does not wish to be bound by the biological theory of theimmuno stimulation, actual immunostimulatory efficacy of the compoundscan be demonstrated in vivo by assaying splenic size in response toadminstration of the test compound to laboratory test species rats.Generally speaking any compound that exhibits splenic enlargementfollowing dosing of 200 mg/kg or less may be considered animmunostimulant.

Modes of Administration:

The compounds of the invention may be administered at pharmaceuticallyeffective dosages. Such dosages are normally the minimum dose necessaryto achieve the desired therapeutic effect; in the treatment of chromicpain, this amount would be roughly that necessary to reduce thediscomfort caused by the pain to tolerable levels.

For human adults such doses generally will be in the range 0.1-5000mg/day; more preferably in the range 1 to 3000 mg/day, still morepreferably in the range of 10 mg to 1000 mg/day. However, the actualamount of the compound to be administered in any given case will bedetermined by a physician taking into account the relevantcircumstances, such as the severity of the pain, the age and weight ofthe patient, the patient's general physical condition, the cause of thepain, and the route of administration.

The compounds are useful in the treatment of pain in a mammal;particularly a human being. Preferably, the patient will be given thecompound orally in any acceptable form, such as a tablet, liquid,capsule, powder and the like. However, other routes may be desirable ornecessary, particularly if the patient suffers from nausea. Such otherroutes may include, without exception, transdermal, intraperitonial,parenteral, subcutaneous, intranasal, intrathecal, intramuscular,intravenous and intrarectal modes of delivery. Another aspect of theinvention is drawn to therapeutic compositions comprising the novelcompounds of the invention and pharmaceutically acceptable salts ofthese compounds and a pharmaceutically acceptable excipient. Such anexcipient may be a carrier or a diluent; this is usually mixed with theactive compound, or permitted to dilute or enclose the active compound.If a diluent, the carrier may be solid, semi-solid, or liquid materialthat acts as an excipient or vehicle for the active compound. Theformulations may also include wetting agents, emulsifying agents,preserving agents, sweetening agents, and/or flavoring agents. If usedas in an ophthalmic or infusion format, the formulation will usuallycontain one or more salt to influence the osmotic pressure of theformulation.

In another aspect, the invention is directed to methods for thetreatment of pain, particularly chronic pain, through the administrationof one or more of the novel or otherwise known compounds of theinvention, or of pharmaceutically acceptable salts thereof to a mammalin need thereof As indicated above, the compound will usually beformulated in a form consistent with the desired mode of delivery.

Compounds of the invention which are immuno stimulants are administeredsubject to the same basic principles as the compounds having analgesicactivity, in doses which are best determined on a case-by-case and/orspecies-by-species and, in case of humans, at times on apatient-by-patient basis. Generally speaking the effective dose will bein the range of 10 μg/kg to 200 mg/kg.

Synthetic Methods for Obtaining the Compounds of the Invention,Experimental

The compound of the invention can be synthesized by utilizing thesynthetic methods described in the experimental below, or suchmodifications of the below described experimental methods which willbecome readily apparent to those skilled in the art in light of thepresent disclosure.

General

¹H NMR spectra were recorded at ambient temperature with an Avance 300(Bruker) spectrometer. The compounds were analyzed by reverse phase highperformance liquid chromatography (HPLC) using a Waters AutopurificationSystem equipped with a Waters 2525 Pump, a Waters 2696 photodiode arraydetector, and a XTerra column (Part. No. 186000482, 5 μm, C18, 4.5×50mm).

The HPLC method used was a gradient of 5% solvent B to 100% in 7 min.Solvent A was H₂O with 0.05% TFA and solvent B was CH₃CN with 0.05% TFA(Method A).

Melting points were measured with a Büchi B-545 melting point apparatusand were uncorrected. To isolate reaction products the solvent wereremoved by evaporation using a vacuum rotatory evaporator, the waterbath temperature not exceeding 40° C.

General Synthetic Routes

The compound of the invention can be synthesized by utilizing thesynthetic methods described in a general sense immediately below and inmore detail in the experimental section of the present application, orby such modifications of the below described general and experimentalmethods which will become readily apparent to those skilled in the artin light of the present disclosure.

A general synthetic route to the compound of the present invention whichare substituted “1-hydroxyl-propyl amines” may lead through thesynthesis of the corresponding substituted “3-hydroxyl-propyl amide”compounds, followed by reduction of the carbonyl group of the“carboxylic acid amide” moiety with a reducing agent such as lithiumaluminum hydride, or like reducing agent.

This reaction is illustrated in General Synthetic Scheme A, where,generally speaking, the variables have the meaning described in theSummary Section of the present application for patent. A person ofordinary skill in the art of organic synthesis will nevertheless readilyunderstand that depending on the nature of the substituents designatedR₁, R₂ and R₁₀ certain groups may need to be protected for theperformance of the reduction step.

The substituted “3-hydroxyl-propyl amide” compounds can, generallyspeaking, be synthesized as described below in the following GeneralReaction Scheme 1 and General Reaction Scheme 2.

Thus, in accordance with General Scheme 1, methyl isocyanoacetate (orethyl isocyanoacetate available commercially) is reacted with an “amine”which includes the R₁ and R₂ groups to provide the 2-isocyanoacetic acidamide derivative shown in

General Reaction Scheme 1. Typical examples for the amines used in thereaction are pyrrolidine, piperidine, azetidine, morpholine,2,5-dihydro-1H-pyrrole, dialkylamines such as diethylamine, 3-fluoro-,3,3-difluoro or 3-hydroxy substituted pyrrolidines. The 2-isocyanoaceticacid amide derivative is then reacted in methanol in the presence ofbase (such as KOH) with an “aldehyde” which includes the R₁₀ group toprovide a trans “oxazoline” with high diastereoselectivity (trans:cisratios generally >97:3) as shown in General Reaction Scheme 1. The transoxazoline is then treated in methanol with a strong acid, such as HCl,to open the ring and to provide thethreo-3-substituted-3-hydroxy-2-amino-propionic acid amide intermediates(with a threo:erythro ratios generally >97:3) as shown in GeneralReaction Scheme 1.

Compounds of Formula 1 and/or of General Structure 1, where the aminogroup of formula NHR₁R₂ is a weaker nucleophile, such as indoline,thiomorpholine and the like, can be made as illustrated in ReactionScheme 2 for the synthesis of intermediate compounds(±)-threo-2-amino-3-hydroxy-1-(indolin-1-yl)-3-(pyridin-4-yl)propan-1-onedihydrochloride Compound 243 and(±)-threo-2-amino-3-hydroxy-1-(thiazolidin-3-yl)-3-(pyridin-4-yl)propan-1-onedihydrochloride Compound 242.

In Reaction Scheme 2 EDCl stands for1-(3-dimethylaminopropyl)-ethylcarbodiimide hydrochloride; HOBT standsfor 1-hydroxybenzotriazole; BOC₂O stands for di-t-butyl-dicarbonate andTEA stands for triethylamine Compounds 242 and 243 can be reduced, asillustrated in General Synthetic Scheme A to provide compounds of theinvention.

Another general synthetic route may follow in general terms thesynthesis of Compound 1, Compound 2 and Compound 3, specificallydescribed in detail in the experimental section below, modified withsuch modifications which in light of the present disclosure will becomereadily apparent to a person of ordinary skill in the art.

Isomerically pure and/or enantiomerically pure compounds and furtherderivatives of the 3-substituted-3-hydroxy-2-amino-propionic acid amideintermediates or of the substituted 1-hydroxy propylamines of theinvention are obtained by separation techniques and reactions which, perse, are well known to the synthetic chemist. Some of the typicalseparation techniques and reactions are generally described below.

Separation of threo and erythro isomers, when both are formed in thereactions leading to the compounds of the invention, can typically beperformed by chromatographic methods. The chromatographic separation mayoccur the level of the substituted 3-hydroxyl-propionic acid amideintermediate compounds or at the level of the substituted 1-hydroxylpropyl amine compounds of the invention.

The more abundantly formed threo isomers can also be converted into theerythro isomers by oxidizing to the ketone level the hydroxyl group inthe 3 position of the propanoic acid moiety and subsequently reducingthe resulting ketone to the hydroxyl level in the intermediate3-substituted-3-hydroxy-2-amino-propionic acid amide compounds or in thecompounds of the invention.

Separation of enantiomeric mixtures can be performed on Chiralpakcolumns which are well known in the art.

The amino function in the 2-position of the propyl amine moiety is,generally speaking, more reactive towards acylation and carbamoylationthan the hydroxyl group in the 1 position. Therefore, acylatedderivatives of the 2-amino function can be prepared by using acylchlorides such as acetyl chloride and hexanoyl chloride. Or the1-hydroxy and 2-amino groups of the compounds of the invention can beacylated in the same reaction. Carbamate derivatives of the 2-aminofunction can be obtained by using chloroformates, such asbenzylchloroformate. A tertiary butyl carbamoyl function orbenzyl-carbamoyl function can also serve as a removable protecting groupof the 2-amino function.

Alkylation of the 2-amino function can be performed by condensing thecompound bearing the 2-NH₂ group with an aldehyde to obtain a Schiffbase intermediate which can be reduced, without isolation, to providethe N-alkyl, arylalkyl or heteroaryl-alkyl compounds of the invention.

DETAILED DESCRIPTION OF THE SYNTHESIS OF PREFERRED COMPOUNDS(EXPERIMENTAL) Preparation ofD-threo-2-amino-3-morpholino-1-phenylpropan-1-ol dihydrochlorideCompound 4 (R)-Methyl 1-((S)-1-phenylethyl)aziridine-2-carboxylate EBE06044B

To solution of methyl 2,3-dibromopropionate (25 mL, 198 mmol) in tolueneat 5° C. was added triethylamine (55 mL, 0.39 mmol) in toluene (100 mL).After stirring for 5 min (S)-(1)-phenethylamine (25 mL, 198 mmol) intoluene (100 mL) was added dropwise. The suspension was refluxed for 3 hand allowed to cool down, filtered and the volatiles were evaporatedunder reduced pressure to give a residue that was purified by columnchromatography (950 g of silica gel) with a gradient of 0-20% EtOAc incyclohexane to yield to (S)-methyl1-((S)-1-phenylethyl)aziridine-2-carboxylate EBE 06044A as a yellow oil(17.31 g, 43% yield) and (R)-methyl1-((S)-1-phenylethyl)aziridine-2-carboxylate EBE 06044B as a yellow oil(15.14 g, 37% yield).

MW: 205.3; Yield EBE 06044B: 37%; Yellow Oil. Yield: EBE 06044A: 43%,Yellow Oil.

R_(f): EBE 06044A=0.5; R_(f): EBE 06044B=0.35 (EtOAc:cyclohexane=25:75).

¹H-NMR (CDCl₃,) EBE 06044A: 1.47 (d, 3H, J=6.6 Hz, CH₃), 1.60 (d, 1H,J=6.4 Hz, CH), 2.13 (d, 1H, J=2.6 Hz), 2.21 (dd, 1H, J=3.2 Hz, J=6.4Hz), 2.54 (q, 1H, J=6.6 Hz), 3.75 (s, 3H, OCH₃) 7.23-7.40 (m, 5H, ArH).

¹H-NMR (CDCl₃,) EBE 06044B: 1.46 (d, 3H, J=6.6 Hz, CH₃), 1.79 (d, 1H,J=6.6 Hz, CH), 2.08 (d, 1H, J=3.11 Hz, 6.6 Hz), 2.34 (dd, 1H, J=3.1 Hz,J=1.0 Hz), 2.56 (q, 1H, J=6.6 Hz), 3.67 (s, 3H, OCH₃) 7.24-7.36 (m, 5H,ArH).

¹³C-NMR (CDCl₃,) EBE 06044B: 23.5, 35.0, 36.9, 52.2, 69.8, 126.5, 127.2,128.5, 143.6, 171.1.

HPLC: Method A, detection at 254 nm, EBE 06044B RT=6.11 min, peak area92.9%.

((R)-1-((S)-1-Phenylethyl)aziridin-2-yl)methanol EBE 06046

A 250 mL round bottom flask was charged with anhydrous THF (100 mL) andLiAlH₄ (2.77 g, 73.1 mmol). While the suspension is stirred at 0° C., asolution of (S)-methyl 1-((S)-1-phenylethyl)aziridine-2-carboxylate EBE06044B (10.0 g, 48.7 mmol) in THF (50 mL) was added dropwise over 20min. The dropping funnel was washed with THF (2×3 mL) and allowed toreact 20 min at 0° C. Maintaining the reaction mixture at 0° C., asolution of KOH (10%, 20 mL) was added dropwise for 20 min (caution thereaction is exothermic). The mixture was stirred for 0.5 h at 25° C. andthe white precipitate removed by filtration through a celite pad thatwas washed with diethyl ether (30 mL). The combined organic filtrateswere washed with NaH₂PO₄ and the aqueous layer was extracted with Et₂O(3×30 mL). The combined organic phase were dried with Na₂SO₄ andconcentrated to give ((R)-1-((S)-1-phenylethyl)aziridin-2-yl)methanolEBE 06046 as a white solid (10.4 g, 90% yield).

MW: 177.2; Yield: 90%; White Solid; Mp (° C.): 37.7.

¹H-NMR (CDCl₃,): 1.43 (d, 3H, J=6.6 Hz, CH₃), 1.49 (d, 1H, J=6.5 Hz,CH), 1.65-1.71 (m, 1H, CH), 1.92 (d, 1H, J=3.5 Hz, NCH), 2.26 (s, 1H,OH), 2.53 (q, 1H, J=6.6 Hz, NCH), 3.32-3.37 (m, 1H, OCH₂), 3.56 (m, 1H,OCH₂), 7.23-7.35 (m, 5H, ArH).

¹³C-NMR (CDCl₃,): 22.9, 31.4, 39.3, 62.5, 69.4, 126.6, 127.3, 128.6,144.5.

(R)-1-((S)-1-Phenylethyl)aziridine-2-carbaldehyde EBE 06048

A three neck, 250 mL round bottom flask was equipped with a lowtemperature thermometer and two (2) equalizing dropping funnels. One ofthese was connected to a nitrogen line and charged with a solution of((R)-1-((S)-1-phenylethyl)aziridin-2-yl)methanol EBE 06046 (7.0 g, 39 5mmol) in CH₂Cl₂ (75 mL), the other was charged with a solution of DMSO(9.25 g, 118 5 mmol) in CH₂Cl₂ (11 mL). To a solution of oxalyl chloride(7.5 g, 59 3 mmol) in CH₂Cl₂ (90 mL) under N₂ at −78° C., the DMSOsolution was added dropwise during 20 min and stirred for 20 min. EBE06046 (7.0 g, 39 5 mmol) in CH₂Cl₂ (75 mL) was added dropwise over 50min. then the dropping funnel was charged with DIEA (42.6 mL, 237 mmol)in CH₂Cl₂ (10 mL) and the reaction mixture was stirred for 30 min at−45° C. The DIEA solution was added over 5 min with the reaction mixtureat −78° C. and the reaction was allowed to warm to room temperature. Thereaction mixture was washed with H₂O (3×50 mL), dried over MgSO₄,filtered, evaporated. The crude product obtained was purified by columnchromatography on silica with a gradient of 0-20% [v/v] EtOAc incyclohexane to give (R)-1-((S)-1-phenylethyl)aziridine-2-carbaldehydeEBE 06048 as a yellow oil (5.59 g, 81% yield).

MW: 175.2; Yield: 81%; Yellow Oil.

R_(f): EBE 06048: 0.3 (EtOAc:cyclohexane=20:80).

¹H-NMR (CDCl₃,): 1.47 (d, 3H, J=6.6 Hz, CH₃), 1.94 (d, 1H, J=6.7 Hz,NCH₂), 2.08 (dt, J=2.9 Hz, J=6.4 Hz, NCH), 2.37 (d, 1H, J=2.6 Hz, NCH₂),2.61 (q, 1H, J=6.6 Hz, NCH), 7.20-7.38 (m, 5H, ArH), 8.92 (d, 1H, J=6.2Hz).

¹³C-NMR (CDCl₃,): 22.7, 32.1, 43.2, 68.1, 125.5, 126.5, 127.6, 142.4,198.7.

(R)-Phenyl((R)-1-((S)-1-phenylethyl)aziridin-2-yl)methanolEBE 06066

To a solution of bromobenzene (4.93 g, 31 4 mmol) in THF 125 mL undernitrogen at −78° C. was added t-BuLi (1.7 M in pentane, 50 mL). Themixture was stirred for 0.5 h at room temperature. The mixture wascooled down to −78° C. and a solution of(R)-1-((S)-1-phenylethyl)aziridine-2-carbaldehyde EBE 06048 (2.5 g, 14.3mmol) in THF (16.7 mL) at −78° C. was added dropwise. The reactionmixture was treated with H₂O (20 mL), the organic layer was separatedand the aqueous phase was extracted with EtOAc. The combined organiclayers were dried over MgSO₄, filtered and concentrated in vacuo to givea residue that was purified by column chromatography using a gradient of0-20% [v/v] EtOAc in cyclohexane to give(R)-phenyl((R)-1-((S)-1-phenylethyl)aziridin-2-yl)methanol EBE 06066(3.13 g, 86% yield).

MW: 253.3; Yield: 86%.

R_(f):=0.3 (EtOAc:cyclohexane=20:80).

¹H-NMR (CDCl₃,): 1.47 (d, 3H, J=6.6 Hz, CH₃), 1.57 (d, 1H, J=6.5 Hz,CH), 1.79 (dt, 1H, J=3.5 Hz, J=8.7 Hz, CH), 2.04 (d, 1H, J=3.5 Hz, OCH),2.35 (bs, 1H, OH), 2.53 (q, 1H, J=6.5 Hz, CH), 4.23 (d, 1H, J=5.7 Hz,OCH), 7.07-7.13 (m, 2H, ArH), 7.16-7.20 (m, 3H, ArH), 7.24-7.34 (m, 5H,ArH).

¹³C-NMR (CDCl₃,): 22.4, 32.0, 44.6, 69.4, 74.1, 125.8(2×C), 126.9 (2×C),127.3, 127.6, 128.2 (2×C), 128.7 (2×C), 142.0, 144.2.

[ ]²² _(D)=−71.53 (c=0.59, CHCl₃).

D-threo-2-((S)-1-Phenylethylamino)-3-morpholino-1-phenylpropan-1-oldihydrochloride Compound 5

To a solution of(R)-phenyl((R)-1-((S)-1-phenylethyl)aziridin-2-yl)methanol EBE 06066(1.5 g, 5.92 mmol) in CH₃CN (19 mL) at RT was added iodotrimethylsilane(3.55 g, 17.8 mmol). The solution was stirred for 2 h and morpholine(1.032 g, 11.84 mmol) was added. After 2 h at reflux, the reactionmixture was treated with HCl (1M) to reach pH=1 and stirred for 10 min.After a slow addition of NaHCO₃ to reach pH=9, the product was extractedwith EtOAc, dried over Na₂SO₄, filtered to give after evaporation acrude brown oil that was purified by column chromatography using agradient of 0-20% [v/v] MeOH in EtOAc to giveD-threo-2-((S)-1-phenylethylamino)-3-morpholino-1-phenylpropan-1-ol EBE06068A (0.831 g, 42%) as a pale brown solid. To a solution ofD-threo-2-((S)-1-phenylethylamino)-3-morpholino-1-phenylpropan-1-ol EBE06068A (0.100 g, 0.294 mmol) in ethanol (1 mL) was added a solution ofHCl (0.8 M, 0.816 mL) in EtOH. Evaporation of the volatiles afforded toD-threo-2-((S)-1-phenylethylamino)-3-morpholino-1-phenylpropan-1-oldihydrochloride Compound 5 as white solid (0.125 g, 100%).

MW: 412.37; Yield: 42%; White Solid; Mp (° C.): 157.2 (dec).

R_(f):0.3 (MeOH:EtOAc=20:80) EBE 06068A.

¹H-NMR (CD₃OD,):1.19 (t, 2H, J=7.0 Hz, NCH₂), 1.71 (d, 3H, J=6.8 Hz,CH₃), 3.45 (m, 2H, J=7.1 Hz, NCH₂), 3.62 (q, 2H, J=7.1 Hz, N—CH₂), 3.97(t, 4H, J=4.5 Hz, OCH₂), 4.06 (m, 1H, CH—N), 4.75 (q, 1H, J=6.8 Hz,CH—N), 5.21 (d, 1H, J=5.1 Hz, CH—O), 7.44-7.56 (m, 10H, ArH).

MS-ESI m/z (% rel. Int.): 341.1 ([MH]⁺, 20).

¹³C-NMR (CD₃OD,): 24.4, 54.5 (2×C), 55.5, 55.9, 60.0, 67.0 (2×C), 75.6,126.3 (2×C), 126.5 (2×C), 127.0, 127.1, 128.1 (2×C), 128.5 (2×C), 142.2,145.3.

HPLC: Method A, detection at 254 nm, Compound 5 RT=4.41 min, peak area99%.

Threo-2-Amino-3-morpholino-1-phenylpropan-1-ol dihydrochloride Compound4

To a solution ofD-threo-2-((S)-1-phenylethylamino)-3-morpholino-1-phenylpropan-1-ol EBE06068A (0.400 g, 1.17 mmol) in MeOH (6 mL) at RT was added acetic acid(0.133 mL, 2.35 mmol). The reaction vessel was flushed with nitrogen andPd(OH)₂ (25% weight, 0.150 g) was added. The nitrogen atmosphere wasexchanged with hydrogen using three cycle of vacuum and hydrogenaddition using a balloon of hydrogen. After stirring for 16 h underhydrogen the reaction mixture was filtrated through celite to give EBE06070A the acetate salt of(2R)-amino-3-morpholin-4-yl-(1R)-phenyl-propan-1-ol (0.279 g, 98%yield). To as solution of EBE 06070A the acetate salt of(2R)-amino-3-morpholin-4-yl-(1R)-phenyl-propan-1-ol (0.100 g, 0.338mmol) in ethanol (1 mL) was added a solution of HCl (0.8 M, 0.930 mL) inEtOH. Evaporation of the volatiles afforded toD-threo-2-amino-3-morpholino-1-phenylpropan-1-ol dihydrochlorideCompound 4 (0.104 g, 100% yield) as an off white solid. (Adapted fromShin, S-H.; Han, E. Y.; Park, C. S.; Lee, W. K.; Ha, H.-J. TetrahedronAsymmetry, 2000, 11, 3293-3301).

MW: 309.23; Yield: 99%; Off White Solid; Mp (° C.): 183.4.

¹H-NMR (CD₃OD,): 3.30-3.77 (m, 6H, CH₂N), 3.92-4.05 (m, 4H, CH₂O),4.05-4.16 (m, 1H, CH), 4.85-4.98 (m, 1H, CH), 7.35-7.60 (m, 5H, ArH).

¹³C-NMR (CD₃OD,): 53.2, 58.3, 58.5 (2×C), 64.9 (2×C), 72.6, 128.0 (2×C),130.2 (2×C), 140.3.

MS-ESI m/z (% rel. int.): 237.1 (100, [MH]⁺).

HPLC: Isocratic 10% CH₃CN in H₂O (pH 10, [NH₄OH]=5 mM), detection UV 254nm, Compound 4 RT=6.63 min, peak area 97.3%.

[ ]²² _(D)=−10.7 (c=1.00, MeOH).

Preparation of BenzylL-threo-1-hydroxy-3-morpholino-1-phenylpropan-2-ylcarbamatehydrochloride Compound 1 Benzyl(S)-3-hydroxy-1-oxo-1-phenylpropan-2-ylcarbamate TTA 08010B

To a stirred solution of Z-L-Ser-OH (6.00 g, 25.08 mmol) in 32 mL ofanhydrous THF at 0° C. under nitrogen was added dropwise 1 Mphenylmagnesium bromide in THF (32 mL, 200 mmol). (The symbol Zdesignates a benzylcarbamoyl group). The mixture was stirred 15 h at RTunder nitrogen. A solution of 2 M HCl (100 mL) was slowly added at 0° C.and the mixture was partitioned between ethyl acetate (750 mL) andacidic water. The organic layer was washed with water (2×20 mL), 1 Naqueous sodium bicarbonate (2×20 mL), brine (2×20 mL) and dried overMgSO₄. After removing ethyl acetate by evaporation at 30-35° C., thecrude product (4.50 g, 60% yield) was cristallized in a mixture of ethylacetate:hexane=25 mL:20 mL to give benzyl(S)-3-hydroxy-1-oxo-1-phenylpropan-2-ylcarbamate TTA 08010B as a whitesolid (1.40 g, 20% yield).

MW: 299.32; Yield: 20%; White Solid; Mp (° C.): 106.5.

R_(f):0.75 (CH₂Cl₂:MeOH=9:1).

¹H-NMR (CDCl₃,): 2.78 (s, 1H, OH), 3.85-3.93 (m, 1H, CH₂O), 4.00-4.09(m, 1H, CH₂O), 5.14 (s, 2H, ArCH₂O), 5.40 (t, 1H, J=3.3 Hz, CH), 6.17(d, 1H, J=6.4 Hz, NH), 7.35 (s, 5H, ArH), 7.49 (t, 2H, J=7.60 Hz, ArH),7.62 (t, 1H, J=7.1 Hz, ArH), 8.99 (t, 2H, J=7.6 Hz, ArH).

¹³C-NMR (CDCl₃,): 58.3, 64.6, 67.3, 128.1, 128.3, 128.6, 128.7, 129.0,134.1, 136.0, 156.6, 196.6.

MS-ESI m/z (% rel. Int.): 300.1 ([MH]⁺, 5), 256.1 (100).

HPLC: Method A, detection UV 254 nm, TTA 08010B RT=5.40 min, peak area98.5%.

[ ]²² _(D)=−5.8 (c=1.00, MeOH).

Benzyl L-threo-1,3-dihydroxy-1-phenylpropan-2-ylcarbamate TTA 08012

To a stirred solution of benzyl(S)-3-hydroxy-1-oxo-1-phenylpropan-2-ylcarbamate TTA 08010B (1.40 g,4.70 mmol) in 28 mL of anhydrous THF at −78° C. under nitrogen was addedslowly dropwise 1 M DIBAL-H in hexane (18.8 mL, 18.80 mmol). The mixturewas stirred 2 h at −78° C. then 1.5 h at RT. A solution of 2 M HCl (35mL) was slowly added at −20° C. and the mixture was partitioned betweenethyl acetate (750 mL) and acidic water. The organic phase was washedwith water (2×20 mL), brine (2×20 mL) and dried over MgSO₄. Afterremoving ethyl acetate by evaporation at 30-35° C., the crude productwas purified by column chromatography on silica (CH₂Cl₂:MeOH=98:2 to97:3) to give benzyl L-threo-1,3-dihydroxy-1-phenylpropan-2-ylcarbamateTTA 08012 as a white solid (1.10 g, 78% yield).

MW: 301.34; Yield: 78%; White Solid; Mp (° C.): 102.5.

R_(f):0.30 (CH₂Cl₂:MeOH=95/5).

¹H-NMR (CDCl₃,): 3.08 (t, 1H, J=5.0 Hz, OH), 3.59 (d, 1H, J=3.1 Hz, OH),3.64-3.78 (m, 2H, CH₂O), 3.80-3.89 (m, 1H, CH), 4.95 (s, 2H, ArCH₂O),5.57 (d, 1H, J=8.3 Hz, NH), 7.17-7.38 (m, 10H, ArH).

¹³C-NMR (CDCl₃,): 57.5, 63.6, 66.9, 73.8, 126.0, 127.8, 127.9, 128.1,128.5, 128.6, 136.2, 141.0, 156.9.

MS-ESI m/z (% rel. Int.): 302.0 ([MH]⁺, 5); 132.0 (100).

HPLC: Method A, detection UV 254 nm, TTA 08012 RT=5.00 min, peak area99.5%.

[ ]²² _(D)=+39.4 (c=1.00, MeOH).

Benzyl threo-1-hydroxy-3-morpholino-1-phenylpropan-2-ylcarbamatehydrochloride Compound 1

To a stirred solution of benzylL-threo-1,3-dihydroxy-1-phenylpropan-2-ylcarbamate TTA 08012 (1.00 g,3.30 mmol) in 13 mL of pyridine at −10° C. was added dropwisemethanesulfonyl chloride (0.27 mL, 3.50 mmol). The mixture was stirred 6h at 20° C. under nitrogen. Pyridine was removed by evaporation at30-35° C. and the residue was partitioned between ethyl acetate (250 mL)and 0.1 N HCl (20 mL). The organic phase was washed with water (20 mL),brine (20 mL), dried over MgSO₄ and evaporated to give after dryingL-threo-1-hydroxy-3-methanesulfonyl-1-phenylpropan-2-ylcarbamate TTA08014 (1.25 g , 65% yield).

To a stirred solution of crude benzylL-threo-1-hydroxy-3-methanesulfonyl-1-phenylpropan-2-ylcarbamate TTA08014 (1.25 g, 3.30 mmol) in 6 mL of DMF at RT was added morpholine (1.2mL, 13.20 mmol). The mixture was stirred 15 h at 50° C. under nitrogen.DMF was evaporated and the residue was partitioned between ethyl acetate(250 mL) and 1 N aqueous sodium bicarbonate (20 mL). The organic phasewas washed with water (20 mL), brine (20 mL) and dried over MgSO₄. Afterevaporation the crude product was purified by column chromatography onsilica (CH₂Cl₂:MeOH=98:2 to 97:3) to give benzylL-threo-1-hydroxy-3-morpholino-1-phenylpropan-2-ylcarbamate as an oil(380 mg, 31% yield). The hydrochloride salt was obtained from 100 mg ofthe free base in diethylether at 0° C. using a solution 0.3 M HCl indiethylether. The precipitate was filtered and dry to give benzylL-threo-1-hydroxy-3-morpholino-1-phenylpropan-2-ylcarbamatehydrochloride Compound 1 as a white solid (70 mg, 65% yield).

MW: 406.90; Yield: 20%; White Solid; Mp (° C.): 144.5.

R_(f):0.40 (CH₂Cl₂:MeOH=95:5).

¹H-NMR (CD₃OD,): 3.14-3.77 (m, 6H, CH₂N), 3.70-4.07 (m, 4H, CH₂O),4.30-4.33 (m, 1H, CH), 4.90-5.06 (m, 3H, CH, ArCH₂O), 7.20-7.43 (m, 10H,ArH).

¹³C-NMR (CD₃OD,): 51.2, 51.8, 53.2, 59.3, 63.2, 66.3, 72.5, 125.8,127.2, 127.3, 127.5, 127.8, 127.9.

MS-ESI m/z (% rel. Int.): 371.0 ([MH]⁺, 100).

HPLC: Method A, detection UV 254 nm, Compound 1 RT=4.40 min, peak area96.5%.

[ ]²² _(D)=+13.9 (c=1.00, MeOH).

Preparation of threo-2-amino-3-morpholino-1-phenylpropan-1-oldihydrochloride Compound 2

To a stirred solution of benzylL-threo-1-hydroxy-3-morpholino-1-phenylpropan-2-ylcarbamate (Compound 1,0.26 g, 0.70 mmol) in 20 mL of MeOH at RT was added Pd—C 10% (140 mg).The mixture was satured with hydrogen and stirred for 24 h at RT underhydrogen atmosphere (balloon). The catalyst Pd—C 10% was removed byfiltration on celite and the solution was evaporated. The crude productwas purified by column chromatography on silica(CH₂Cl₂:MeOH:NH₄OH=79:20:1 to 75:20:5) to giveL-threo-2-amino-3-morpholino-1-phenylpropan-1-ol as an oil (100 mg, 60%yield). The hydrochloride salt was obtained from 83 mg of the free basein diethylether at 0° C. using 0.3 M HCl in diethylether. Afterprecipitation in diethylether, filtration and dryingL-threo-2-amino-3-morpholino-1-phenylpropan-1-ol dihydrochlorideCompound 2 was obtained as a white solid (80 mg, 74% yield).

MW: 309.23; Yield: 44%; White Solid; Mp (° C.): 166.4-170.9.

R_(f):0.20 (CH₂Cl₂:MeOH=9:1).

¹H-NMR (CD₃OD,): 3.30-3.77 (m, 6H, CH₂N), 3.92-4.05 (m, 4H, CH₂O),4.05-4.16 (m, 1H, CH), 4.85-4.98 (m, 1H, CH), 7.35-7.60 (m, 5H, ArH).

¹³C-NMR (CD₃OD,): 53.1, 54.9, 58.5, 64.8, 72.6, 127.2, 128.0, 130.2,140.3.

MS-ESI m/z (% rel. Int.): 237.0 ([MH]⁺, 100).

HPLC: Method A, detection UV 254 nm, Compound 2 RT=0.90 min, peak area98.0%.

[ ]²² _(D)=+10.8 (c=1.00, MeOH), free base: [ ]²² _(D)=−6.1 (c=0.25,CHCl₃).

Preparation of benzylL-threo-1-acetoxy-3-morpholino-1-phenylpropan-2-ylcarbamatehydrochloride Compound 3 BenzylL-threo-1-acetoxy-3-morpholino-1-phenylpropan-2-ylcarbamatehydrochloride Compound 3

To a stirred solution of benzylL-threo-1-hydroxy-3-morpholino-1-phenylpropan-2-ylcarbamatehydrochloride (Compound 1, 0.510 g, 1.25 mmol) in 30 mL of CHCl₃ at RTwere added slowly triethylamine (700 μL, 5.00 mmol) and acetyl chloride(145 μL, 2.00 mmol). The mixture was stirred 10 h at RT under nitrogenand partitioned between a mixture of ice-water (20 mL) and CH₂Cl₂ (100mL). The organic layer was washed with brine (20 mL) and dried overMgSO₄. After evaporation the crude product was purified by columnchromatography on silica (CH₂Cl₂:MeOH=99.5:0.5 to 98:2) to give benzylL-threo-1-acetoxy-3-morpholino-1-phenylpropan-2-ylcarbamate as an oil(0.420 g, 81% yield).

The hydrochloride salt was obtained from 45 mg of the free base indiethylether at 0° C. using a solution of 0.3 M HCl in diethylether. Theprecipitate was filtered and dry to give benzylL-threo-1-acetoxy-3-morpholino-1-phenylpropan-2-ylcarbamatehydrochloride Compound 3 as a white solid (40 mg, 82% yield).

MW: 448.94; Yield: 66%; White Solid; Mp (° C.): 69.9.

R_(f):0.70 (CH₂Cl₂:MeOH=95:5).

¹H-NMR (CD₃OD,): 2.10 (s, 3H, CH₃), 3.14-3.44 (m, 4H, CH₂N), 3.70-4.00(m, 4H, CH₂O), 4.51-4.53 (m, 1H, CH), 4.90-5.13 (m, 2H, ArCH₂O), 5.89(d, 1H, CH), 7.28-7.48 (m, 10H, ArH).

¹³C-NMR (CD₃OD,): 20.8, 52.0, 52.6, 59.7, 64.6, 68.0, 76.5, 127.7,129.0, 129.2, 129.5, 129.8, 137.9, 158.7, 171.3.

MS-ESI m/z (% rel. Int.): 413.0 ([MH]⁺, 100).

HPLC: Method A, detection UV 254 nm, Compound 3 RT=4.70 min, peak area98.5%.

Preparation ofDL-threo-2-(Decanamido)-1-(4-methoxyphenyl)-3-(pyrrolidin-1-yl)propyldecanoate Compound 10 2-Isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098

To stirred and cooled (0° C.) methyl isocyanoacetate (96% technicalgrade, 5.0 g, 47.8 mmol) was slowly added in 0.75 h pyrrolidine (6.5 mL,78 mmol). The mixture was stirred for 1.5 h with continued cooling andthen concentrated. The resulting oil was co-evaporated twice fromCH₂Cl₂:hexane to remove residual pyrrolidine.2-Isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098 was obtained as ayellow solid (6.85 g, 98% yield) and used in the next step withoutpurification.

MW: 138.17; Yield: 98%; yellow solid; Mp (° C.)=73.9.

¹H-NMR (CDCl₃,): 1.81-2.08 (m, 4H, 2×CH₂), 3.35-3.45 (m, 2H, —NCH₂),3.50-3.60 (m, 2H, —NCH₂), 4.23 (s, 2H, CH₂CO).

Trans-(4,5-dihydro-5-(pyridin-3-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04110Btrans-(4,5-Dihydro-5-(4-methoxyphenyl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneSLA 07074

To a stirred and cooled (0° C.) solution of potassium hydroxide (0.37 g,6.57 mmol) in methanol (30 mL) was added a mixture of4-methoxy-benzaldehyde (0.88 mL, 7.23 mmol) and2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098 (1.0 g, 6.57 mmol). Thesolution was stirred 4 h with continued cooling and then concentrated.The residue was partitioned between ethyl acetate and water. The organiclayer was combined with additional ethyl acetate extracts, washed withaqueous sodium chloride and dried over MgSO₄. Concentration afforded acrude product as a glassy solid. Flash chromatography over silica (ethylacetate) yielded totrans-(4,5-dihydro-5-(4-methoxyphenyl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneSLA 07074 as a pale yellow solid (1.2 g, 90.5%).

MW: 274.32; Yield: 90.5%; pale yellow solid; Mp (° C.): 91.2.

R_(f):0.30 (EtOAc).

¹H-NMR (CDCl₃,): 1.75-2.08 (m, 4H, 2×CH₂), 3.40-3.58 (m, 3H, CH₂N), 3.52(s, 3H, CH₃O), 3.88-3.98 (m, 1H, CH₂N), 4.59 (dd, 1H, J=7.6 Hz, J=2.2Hz, CH—N), 6.06 (d, 1H, J=7.6 Hz, CH—O), 6.90 (d, 2H, J=8.7 Hz, ArH),7.01 (d, 1H, J=2.2 Hz, CH═N), 7.25 (d, 2H, J=8.7 Hz, ArH).

MS-ESI m/z (% rel. Int.): 275.1 ([MH]⁺, 10), 247.1 (100).

HPLC: Method A, detection UV 280 nm, SLA 07074 RT=5.2 min, peak area92%.

DL-threo-2-Amino-3-hydroxy-3-(4-methoxyphenyl)-1-(pyrrolidin-1-yl)propan-1-onehydrochloride SLA 07078

To a stirred solution oftrans-(4,5-dihydro-5-(4-methoxyphenyl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneSLA 07074 (1.61 g, 5.93 mmol) in methanol (13 mL) was added hydrochloricacid (1 mL). After heating at 50° C. for 3 h the mixture reaction wasconcentrated and the resulting yellow oil was co-evaporated twice withethyl acetate before solidifying. Trituration (ethyl acetate) and dryingaffordedDL-threo-2-amino-3-hydroxy-3-(4-methoxyphenyl)-1-(pyrrolidin-1-yl)propan-1-onehydrochloride SLA 07078 as a white solid (1.64 g, 93%).

MW: 300.78; Yield: 93%; white Solid; Mp (° C.): 177.0.

¹H-NMR (CD₃OD,): 1.32-1.50 (m, 1H, CH₂), 1.50-1.88 (m, 3H, CH₂),2.15-2.28 (m, 1H, CH₂N), 3.15-3.42 (m, 4H, 2×CH₂N), 3.79 (s, 3H, CH₃O),4.06 (d, 1H, J=9.2 Hz, CH—N), 4.78 (d, 1H, J=9.2 Hz, CHO), 6.94 (d, 2H,J=8.5 Hz, ArH), 7.34 (d, 2H, J=8.5 Hz, ArH).

¹³C-NMR (CD₃OD,): 24.8, 26.6, 47.2, 47.6, 55.9, 59.6, 73.9, 115.0 (2×C),128.9 (2×C), 132.5, 161.7, 166.4.

DL-threo-2-Amino-1-(4-methoxyphenyl)-3-(pyrrolidin-1-yl)propan-1-olCompound 9

To a stirred suspension ofDL-threo-[5-(4-methoxy-phenyl)-4,5-dihydro-oxazol-4-yl]-pyrrolidin-1-yl-methanoneSLA 07078 (1.61 g, 5.35 mmol) in tetrahydrofuran (200 mL) under nitrogenatmosphere was slowly added, in two portions, lithium aluminum hydride(1.22 g, 32.12 mmol) at 0° C. The mixture reaction was stirred at RT for17 h, and then quenched by a slow, dropwise addition of water (50 mL).The white suspension was then concentrated to remove THF and taken backup in a mixture of 300 mL CH₂Cl₂ and 1N aqueous hydrochloric acid (50mL). The aqueous layer was basified to pH=10-11 by a slow addition of 1Naqueous sodium hydroxyde. The organic layer was removed, combined withadditional CH₂Cl₂ extracts (4×200 mL) and dried over MgSO₄, filtered andevaporated. The crude product was purified by column chromatography onsilica (CH₂Cl₂:MeOH:NH₃=94:05:01). After evaporation and drying,DL-threo-2-amino-1-(4-methoxyphenyl)-3-(pyrrolidin-1-yl)propan-1-olCompound 9 was obtained (0.62 g, 46%) as a pale yellow solid.

MW: 250.34; Yield: 46%; Pale Yellow Solid; Mp (° C.): 77.7.

R_(f):0.35 (CH₂Cl₂:MeOH:NH₃=94:05:01).

¹H-NMR (CDCl₃,): 1.65-1.87 (s, 4H, 2×CH₂), 2.40-2.90 (m, 9H, CH₂N, NH₂ &OH), 3.11-3.17 (m, 1H, CH—N), 3.81 (s, 3H, CH₃O), 4.61 (d, 1H, J=3.8 Hz,CH—O), 7.89 (d, 2H, J=8.6 Hz, ArH), 7.26 (d, 2H, J=8.5 Hz, ArH).

¹³C-NMR (CDCl₃,): 23.6 (2×C), 54.5, 54.7 (2×C), 55.3, 60.1, 75.9, 113.6,127.4, 134.4, 158.8.

MS-ESI m/z (% rel. Int.): 251.1 ([MH]⁺, 100).

DL-threo-2-(Decanamido)-1-(4-methoxyphenyl)-3-(pyrrolidin-1-yl)propyldecanoate Compound 10

To a stirred solution ofDL-threo-2-amino-1-(4-methoxy-phenyl)-3-pyrrolidin-1-yl-propan-1-olCompound 9 (0.15 g, 0.60 mmol) in dichloromethane (10 mL) were addedN-hydroxysuccinimide (0.07 g, 0.60 mmol), triethylamine (0.10 mL, 0.63mmol) and decanoyl chloride (112 μL, 0.54 mmol) under nitrogenatmosphere. The mixture reaction was stirred at RT for 22 h andpartitioned between methylene chloride and 1 N aqueous sodium hydroxide.The organic layer was dried over MgSO₄, filtered and evaporated. Thecrude product was purified by column chromatography on silica(CH₂Cl₂:MeOH=95:05).DL-threo-2-(Decanamido)-1-(4-methoxyphenyl)-3-(pyrrolidin-1-yl)propyldecanoate Compound 10 was obtained as a white oil (0.104 g, 31%).

MW: 558.84; Yield: 40%; White Oil.

R_(f):0.35 (CH₂Cl₂:MeOH=95:05).

¹H-NMR (CDCl₃,): 0.88 (t, 6H, J=0.7 Hz, 2×CH₃), 1.26 (s, 14H, 7×CH₂),1.57-1.59 (m, 4H, 2×CH₂), 1.80 (m, 4H, 2×CH₂), 2.10-2.50 (m, 5H, CH₂),2.65-2.76 (m, 5H, CH₂), 3.79 (s, 3H, CH₃O), 4.54 (m, 1H, CH—N), 5.89 (d,1H, J=6.2 Hz, CH—O), 6.16 (d broad, 1H, J=8.8 Hz, NH), 6.85 (d, 2H,J=8.7 Hz, ArH), 7.24 (d, 2H, J=8.7 Hz, ArH).

MS-ESI m/z (% rel. int.): 559.5 ([MH]⁺, 100).

HPLC: Method A, detection UV 280 nm, Compound 10 RT=6.99 min, peak area96.4%.

N-(DL-threo-1-Hydroxy-1-(4-methoxyphenyl)-3-(pyrrolidin-1-yl)propan-2-yl)palmitamideor DL-threo-4-MeO-P4 Compound 11

To a stirred solution ofDL-threo-2-amino-1-(4-methoxyphenyl)-3-(pyrrolidin-1-yl)propan-1-olCompound 9 (015 g, 0.60 mmol) in dichloromethane (10 mL) weresuccessively added N-hydroxysuccinimide (0.07 g, 0.60 mmol),triethylamine (0.100 mL, 0.63 mmol) and palmitoyl chloride (0.15 g, 0.54mmol) under nitrogen atmosphere. The mixture reaction was stirred at RTfor 17 h and partitioned between methylene chloride and 1N aqueoussodium hydroxide. The organic layer was dried over MgSO₄, filtered andevaporated. The crude product was purified by column chromatography onsilica (CH₂Cl₂:MeOH=95:05).N-(DL-threo-1-Hydroxy-1-(4-methoxyphenyl)-3-(pyrrolidin-1-yl)propan-2-yl)palmitamideCompound 11 was obtained as a white solid (0.117 g, 40%).

MW: 488.75; Yield: 40%; White Solid; Mp (° C.): 82.3.

R_(f):0.35 (CH₂Cl₂:MeOH=95:05).

¹H-NMR (CDCl₃:0.88 (t, 3H, J=7.0 Hz, CH₃), 1.22-1.33 (m, 16H, 8×CH₂),1.47-1.54 (m, 2H, CH₂), 1.81 (m, 4H, 2×CH₂), 2.09 (t, 2H, J=7.0 Hz,COCH₂), 2.60-2.80 (m, 4H, 2×CH₂), 2.84 (d, 2H, J=5.1 Hz, CH₂), 3.80 (s,3H, CH₃O), 4.23 (m, 1H, CH—N), 5.00 (d, 1H, J=2.2 Hz, CH—O), 5.90 (d,1H, J=7.4 Hz, NH), 6.87 (d, 2H, J=8.7 Hz, ArH), 7.24 (d, 2H, J=8.7 Hz,ArH).

¹³C-NMR (CDCl₃,): 14.1, 22.7, 23.6, 25.6, 29.1, 29.3, 29.4, 29.5, 29.7,29.7, 31.9, 36.8, 52.3, 55.2, 57.8, 75.4, 113.7 (2×C), 127.0 (2×C),133.1, 158.9, 173.6.

MS-ESI m/z (rel. int.): 489.2 ([MH]⁺, 100).

HP LC: Method A, detection UV 280 nm, Compound 11 RT=6.55 min, peak area96.4%.

DL-threo-2-Amino-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-(pyrrolidin-1-yl)propan-1-olCompound 6trans-(4,5-Dihydro-5-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04100

To a stirred and cooled (0° C.) solution of potassium hydroxide (0.43mg, 7.60 mmol) in MeOH (6.5 mL) were added successively1,4-benzodioxan-6-carboxaldehyde (1.31 g, 7.96 mmol) and2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098 (1.0 g, 6.57 mmol). Thesolution was stirred 3 h at 0° C. and then concentrated. The residue waspartitioned between EtOAc (100 mL) and water. The organic layer wascombined with 2 additional EtOAc extracts (2×100 mL), washed with brine,dried over MgSO₄, filtered and evaporated. Concentration afford to acrude product which was purified by column chromatography on silica(EtOAc) to yield, after evaporation and drying, totrans-4,5-dihydro-5-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04100 as a colourless oil (1.76 g, 89% yield).

MW: 440.49; Yield: 89%; colourless oil.

¹H-NMR (CDCl₃,): 1.75-2.10 (m, 4H, 2×CH₂), 3.40-3.59 (m, 6H, 3×CH₂N),3.85-4.00 (m, 1H, CHN), 4.26 (s, 4H, CH₂O), 4.59 (dd, 1H, J=7.5 Hz,J=2.2 Hz, CH—N), 6.00 (d, 1H, J=7.5 Hz, CH—O), 6.75-6.90 (m, 3H, ArH),7.00 (d, 1H, J=2.2 Hz, CH═N).

DL-threo-2-amino-3-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 12

To a stirred solution oftrans-4,5-dihydro-5-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04100 (1.74 g, 5.77 mmol) in methanol (15 mL) was added hydrochloricacid (1 mL). After heating at 50° C. for 3 h the mixture reaction wasconcentrated and the resulting yellow oil was co-evaporated twice withethyl acetate before solidifying. Trituration (ethyl acetate) and dryingaffordedDL-threo-2-amino-3-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 12 as a white solid (1.85 g, 95%).

MW: 328.79; Yield: 95.0%; White Solid; Mp (° C.): 176.2.

¹H-NMR (CD₃OD,): 1.42-1.58 (m, 1H, CH₂), 1.58-1.70 (m, 1H, CH₂),1.70-1.88 (m, 2H, CH₂), 3.20-3.45(m, 4H, N—CH₂), 4.06 (d, 1H, J=9.1 Hz,CH—N), 4.25 (s, 2H, CH₂), 4.75 (d, 1H, J=9.2 Hz, CH—O), 4.89 (s, 2H,CH₂), 6.82-6.95 (m, 3H, ArH).

¹³C-NMR (CD₃OD,): 24.9, 26.7, 47.3, 47.6, 59.5, 65.7, 73.6, 116.4,118.3, 120.3, 133.7, 145.1, 145.6, 166.4.

DL-threo-2-Amino-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-(pyrrolidin-1-yl)propan-1-olCompound 6

To a stirred suspension oftrans-(4,5-dihydro-5-(4-methoxyphenyl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneSLA 07080 (1.79 g, 5.44 mmol) in THF (220 mL) was slowly added at 0° C.,in two portions, LiAlH₄ (1.28 g, 33.7 mmol). The mixture was stirred atRT for 3.5 h and quenched by a slow addition of water at 0° C. (350 mL).The white suspension was concentrated to remove THF and taken back in amixture of CH₂Cl₂ (300 mL) and 1 N aqueous HCl (50 mL). The aqueouslayer was basified to pH=10-11 by slow addition of 1 N aqueous NaOH. Theorganic layer was removed; two more extracts were combined and driedover MgSO₄, filtered and evaporated. Concentration afforded to a crudeproduct as a yellow oil. This material was purified by columnchromatography on silica (CH₂Cl₂:MeOH:NH₄OH 20%=94:5:1) to led toDL-threo-2-amino-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-(pyrrolidin-1-yl)propan-1-olCompound 6 (0.705 g, 46.5% yield) as a near colorless gum.

MW: 278.35; Yield: 46.5%; Colorless Gum.

R_(f):0.20 (CH₂Cl₂:MeOH:NH₄OH 20%=94:5:1).

¹H-NMR (CDCl₃,): 1.70-1.85 (m, 4H, 2×CH₂), 2.40-2.70 (m, 6H, 3×CH₂N—),3.05-3.15 (m, 1H, CH—N), 4.25 (s, 4H, CH₂O), 4.55 (d, 1H, J=2.2 Hz,CH—O), 5.30 (s, 1H, —OH), 6.75-6.90 (m, 3H, ArH).

N-(DL-threo-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)decanamideCompound 7

To a stirred solution ofDL-threo-2-amino-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-(pyrrolidin-1-yl)propan-1-olBLE 04104 (0.186 g, 0.67 mmol) in 10 mL CH₂Cl₂ were added, in order,N-hydroxysuccinimide (0.081 g, 0.70 mmol) in 2 mL CH₂Cl₂, triethylamine(112 μL, 0.80 mmol) and decanoyl chloride (125 μL, 0.60 mmol). Themixture was stirred overnight at RT and then partitioned between CH₂Cl₂and 1 N aqueous sodium hydroxide. The organic layer was dried overMgSO₄, filtered and evaporated and the residue obtained was purified bycolumn chromatography on silica (CH₂Cl₂:MeOH=95:5). A white solidN-(DL-threo-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)palmitamideCompound 7 was obtained (126 mg, 43.5% yield).

MW: 516.76; Yield: 43.5%; White Solid; Mp (° C.): 84.6.

R_(f):0.40 (MeOH:CH₂Cl₂=10:90).

¹H-NMR (CDCl₃,): 0.88 (t, 3H, J=6.7 Hz, CH₃), 1.12-1.39 (m, 12 H),1.40-1.60 (m, 2H, CH₂), 1.72-1.90 (m, 4H, 2×CH₂), 2.10 (t, 2H, J=6.7 Hz,CH₂), 2.55-2.90 (m, 6H), 4.13-4.30 (m, 1H, CH—N), 4.24 (s, 4H, CH₂N),4.91 (d, 1H, J=3.3 Hz, CH—O), 5.90 (d, 1H, J=7.4 Hz, NH), 6.75-6.88 (m,3H, ArH), OH not seen.

¹³C-NMR (CDCl₃,): 14.1, 22.7, 23.6 (2×C), 25.6, 29.1, 29.3, 31.9, 36.8,52.3, 55.1 (2×C), 57.7, 64.3 (2×C), 75.2, 77.2, 115.0, 117.0, 118.9,134.4, 142.8, 143.4, 173.5, 174.8.

MS-ESI m/z (% rel. Int.): 433.1 ([MH]⁺, 100).

HPLC: Method A, detection UV 280 nm, Compound 7, RT =5.2 min, peak area96.2%.

N-(DL-threo-1-(2,3-Dihydrobenzo[b][1,4]dioxin-6-yl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)palmitamideCompound 8

To a stirred solution ofDL-threo-2-amino-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-(pyrrolidin-1-yl)propan-1-olBLE 04104 (0.158 g, 0.57 mmol) in 10 mL CH₂Cl₂ were added, in order,N-hydroxysuccinimide (0.068 g, 0.59 mmol) in 2 ml CH₂Cl₂, triethylamine(95 μL, 0.68 mmol) and palmitoyl chloride (155 μL, 0.511 mmol) in 3 mLCH₂Cl₂. The mixture was stirred overnight at RT and then partitionedbetween CH₂Cl₂ and 1 N aqueous sodium hydroxyde. The organic layer waspurified by column chromatography on silica using as eluentCH₂Cl₂:MeOH=95:5. A white solidN-(DL-threo-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-1-hydroxy-3-(pyrrolidin-1-yl)propan-2-yl)palmitamideCompound 8 was obtained (148 mg, 50.4% yield).

MW: 516.7; Yield: 50.4%; White Solid; Mp (° C.): 66.4.

R_(f):0.50 (MeOH:CH₂Cl₂=10:90).

¹H-NMR (CDCl₃,): 0.88 (t, 3H, J=6.7 Hz, CH₃), 1.15-1.35 (m, 24 H),1.45-1.58 (m, 2H, CH₂), 1.75-1.90 (m, 4H, 2×CH₂), 2.10 (t, 2H, J=7.4 Hz,CH₂), 2.61 (s, 1H, OH), 2.52-2.72 (m, 4H), 2.72-2.92 (m, 2H), 4.15-4.22(m, 1H, CH—N), 4.24 (s, 4H, CH₂N), 4.92 (d, 1H, J=3.3 Hz, CH—O), 6.08(d, 1H, J=7.4 Hz, NH), 6.75-6.90 (m, 3H, ArH).

MS-EST m/z (% rel. Int.): 517.2 ([MH]⁺, 100).

HPLC: Method A, detection UV 280 nm, Compound 8 RT=6.60 min, peak area97.2%.

Preparation ofDL-threo-2-Amino-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-1-olCompound 46Trans-(4,5-dihydro-5-(pyridin-3-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04110B

A general method D for oxazolines formation is illustrated by thepreparation of BLE 04110B: To a stirred and cooled (0° C.) solution ofpotassium hydroxide (0.55 g, 9.80 mmol) in methanol (10 mL) were added amixture of 3-pyridine carboxaldehyde (1.03 mL, 10.84 mmol) and2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098 (1.50 g, 10.86 mmol).The solution was stirred 3 h at 0° C. and then concentrated. The residuewas partitioned between ethyl acetate (100 mL) and water. The organiclayer was combined with two additional ethyl acetate extracts (2×100mL), washed with aqueous sodium chloride and dried over MgSO₄, filteredand evaporated. Concentration afforded a crude product which waspurified by column chromatography on silica (CH₂Cl₂:MeOH=98:2) to yieldtotrans-(4,5-dihydro-5-(pyridin-3-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04110B (0.95 g, 39%) as a pale yellow pale solid.

MW: 245.28; Yield: 39%; Yellow Pale Solid; Mp (° C.): 107.0.

¹H-NMR (CDCl₃,): 1.78-2.10 (m, 4H, 2×CH₂), 3.40-3.61 (m, 3H, CH₂N),3.90-4.04 (m, 1H, CH₂N), 4.59 (dd, 1H, J=7.7 Hz, J=2.2 Hz, CH—N), 6.21(d, 1H, J=7.7 Hz, CH—O), 7.04 (d, 1H, J=2.2 Hz, O—CH═N), 7.33 (m, 1H,ArH), 7.64 (m, 1H, ArH), 8.59 (d, 2H, J=2.8 Hz, ArH).

¹³C-NMR (CDCl₃,): 24.2, 26.0, 46.4, 46.6, 75.7, 79.3, 123.7, 133.5,135.3, 147.6, 149.9, 155.2, 166.2.

trans-(4,5-Dihydro-5-(pyridin-4-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneCompound 19

Compound 19 was prepared in accordance with method D usingpyridine-4-carbaldehyde (1.88 mL, 19.76 mmol), KOH (1.01 g, 18.00 mmol)in methanol (18 mL) and 2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098(2.73 g, 19.76 mmol). The residue was partitioned between ethyl acetate(200 mL) and water (150 mL). The organic layer was combined withadditional ethyl acetate extracts (2×150 mL), washed with aqueous sodiumchloride (2×150 mL) and dried over MgSO₄, filtered and evaporated.Trans-(4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneCompound 19 was obtained as a white solid (4.32 g, 98% yield).

MW: 245.28; Yield: 98%; White Solid; Mp (° C.)=69.2.

R_(f):0.65 (MeOH:CH₂Cl₂=10:90).

¹H-NMR (CDCl₃,): 1.78-2.06 (m, 4H, 2×CH₂), 3.44-3.60 (m, 3H, CH₂N),3.90-4.01 (m, 1H, CH₂N), 4.52 (dd, 1H, J=7.9 Hz, J=2.2 Hz, CH—N), 6.19(d, J=7.9 Hz, 1H, CH—O), 7.03 (d, 1H, J=2.2 Hz, N═CH—O), 7.24 (dd, 2H,J=4.5 Hz, J=1.5 Hz, ArH), 8.61 (dd, 2H, J=4.5 Hz, J=1.5 Hz, ArH).

A general method for the acidic hydrolysis of oxazolines (Method E) isillustrated in the preparation of Compound 20 which is a substitutedpropionic acid amide and is made from the oxazoline intermediate BLE04110B which can be prepared in accordance with General Synthetic Scheme1.

DL-threo-2-Amino-3-hydroxy-3-(pyridin-3-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 20

To a solution oftrans-(4,5-dihydro-5-(pyridin-3-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04110B (0.932 g, 3.80 mmol) in methanol (10 mL) was addedhydrochloric acid 37% (1.2 mL). After heating (50° C.) the mixture for2.25 h the reaction mixture was concentrated and the crude product wascoevaporated twice with ethyl acetate. After trituration with ethylacetate, filtration and dryingDL-threo-2-amino-3-hydroxy-3-(pyridin-3-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 20 was obtained as a white solid (1.10 g, 94%yield).

MW: 308.2; Yield: 94%; White Solid; Mp (° C.): 123.4.

¹H-NMR (CD₃OD,): 1.65-2.00 (m, 4H, 2×CH₂), 2.82-3.11 (m, 1H, —CH₂N),3.30-3.57 (m, 2H, CH₂N), 3.57-3.77 (m, 1H, CH₂N), 4.54 (d, 1H, J=5.3 Hz,CH—N), 5.38 (d, 1H, J=5.3 Hz, CH—O), 8.15 (dd, 1H, J=7.6 Hz, J=5.0 Hz,ArH), 8.68 (d, 1H, J=7.6 Hz, ArH), 8.89 (d, 1H, J=7.6 Hz, ArH), 8.96 (s,1H, ArH).

¹³C-NMR (CD₃OD,): 24.9, 26.9, 47.7, 48.2, 58.1, 69.6, 128.7, 141.5,141.6, 143.1, 146.5, 165.4.

DL-threo-2-Amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22

Compound 22 was prepared following method E withtrans-(4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneCompound 19 (0.750 g, 3.07 mmol), hydrochloric acid 37% (1.0 mL) andmethanol (10 mL). After 3.0 h at 50° C. and work-upDL-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22 was obtained as a white solid (0.935 g,99%).

MW: 308.28; Yield: 99%; White Solid; Mp (° C.): 117.0.

¹H-NMR (CD₃OD,): 1.75-2.03 (m, 4H, 2×CH₂), 2.93-3.08 (m, 1H, CHN),3.32-3.75 (m, 3H, 2×CH₂), 4.54 (d, 1H, J=5.9 Hz, CH—N), 5.40 (d, 1H,J=5.9 Hz, CH—O), 8.21 (d, 2H, J=5.8 Hz, ArH), 8.94 (d, 2H, J=5.8 Hz,ArH).

MS-ESI m/z (% rel. int.): 236.1 ([MH]⁺, 17), 219 (25), 148 (100). HPLC:Method A, detection UV 254 nm, Compound 22 RT =0.8 min, peak area 96.3%.

DL-threo-2-Amino-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-1-olCompound 46

To a stirred suspension ofDL-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22 (0.86 g, 2.80 mmol) in tetrahydrofuran (108mL) under nitrogen atmosphere was slowly added, in two portions, lithiumaluminum hydride (0.64 g, 16.82 mmol) at 0° C. The mixture reaction wasstirred at RT for 20 h and quenched by a slow, dropwise addition of 2 Naqueous sodium hydroxyde (8.4 mL, 6 eq). The yellow precipitate wasfiltered. The organic layer was washed by water (80 mL) and the organiclayer was removed and combined with additional ethyl acetate extracts(4×200 mL) and dried over MgSO₄, filtered and evaporated. The crudeproduct was purified by column chromatography on silica(CH₂Cl₂:MeOH:NH₃=94:05:01). After evaporation and dryingDL-threo-2-amino-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-1-olCompound 46 was obtained (0.075 g, 12%) as a pale yellow solid.

MW: 221.30; Yield: 12%; Pale Yellow Solid.

R_(f):0.35 (CH₂Cl₂:MeOH:NH₃=90:08:02).

¹H-NMR (CD₃OD,): 1.60-1.80 (m, 4H, 2×CH₂), 2.30-2.80 (m, 6H, 3×CH₂N),3.14-3.19 (m, 1H, CH—NH₂), 4.68 (d, 1H, J=3.0 Hz, CH—O), 7.30 (d, 2H,J=6.0 Hz, ArH), 8.55 (d, 2H, J=6.0 Hz, ArH).

¹³C-NMR (CD₃OD,): 23.5 (2×C), 54.1, 54.7 (2×C), 60.1, 74.5, 121.4 (2×C),149.5 (2×C), 152.1.

MS-ESI m/z (rel. int.): 222.1 ([MH]⁺, 100), 205.0 (80), 189.0 (45),151.0 (70), 134.0 (42), 121.9 (100), 107.9 (40).

1. A compound of the formula

where R₁ is H or alkyl of 1 to 6 carbons, R₂ is H, alkyl of 1 to 6carbons or the R₁ and R₂ groups together with the nitrogen form asaturated or unsaturated 4, 5, 6 or 7 membered ring that optionallyincludes one or two heteroatoms independently selected from N, O and S,said 4, 5, 6 or 7 membered ring optionally being substituted with ahalogen or with an alkyl group having 1 to 6 alkyl groups; R₃ isindependently selected from H, alkyl of 1 to 20 carbons, aryl orheteroaryl, aryl-alkyl or heteroaryl-alkyl where the alkyl moiety has 1to 4 carbons, cycloalkyl of 3 to 6 carbons, said aryl or heteroarylgroups being optionally substituted with 1 to 3 groups independentlyselected from the group consisting of halogen, alkyl of 1 to 6 carbons,alkoxy of 1 to 6 carbons and thioxy of 1 to 6 carbons, or R₃ is CO—R₇ orCO—O—R₇ where R₇ is H, alkyl of 1 to 20 carbons, benzyl, alkyl of 1 to20 carbons substituted with and NH₂ group, with a NHCOOalkyl or with anNH—COalkyl group where the alkyl group has 1 to 6 carbons, or R₇ isaryl, heteroaryl, aryl-alkyl or heteroaryl-alkyl where the alkyl moietyis branched or unbranched and has 1 to 4 carbons, said aryl orheteroaryl groups being optionally substituted with 1 to 3 groupsindependently selected from the group consisting of halogen, alkyl of 1to 6 carbons, alkoxy of 1 to 6 carbons and thioxy of 1 to 6 carbons; R₄is H, alkyl of 1 to 6 carbons or CO—R₈ where R₈ is alkyl of 1 to 6carbons; the wavy lines represent bonds connected to carbons having R orS configuration, and R₁₀ is selected from the groups of formulas (i) and(ii)

where the * indicates the carbon atom to which the remaining moiety ofthe molecule is attached; R₅ and R₆ independently are H, alkyl of 1 to 6carbons, halogen, alkoxy of 1 to 6 carbons or the R₅ and R₆ groupstogether with the atoms to which they are attached jointly form acarbocyclic or a heterocyclic ring, the carbocyclic ring having 5 or 6atoms in the ring, the heterocyclic ring having 5 or 6 atoms in the ringand 1 to 3 heteroatoms independently selected from N, O and S, and saidcarbocyclic or heterocyclic ring jointly formed by R₅ and R₆ beingoptionally substituted with 1 to 6 R₉ groups where R₉ is independentlyselected from halogen, alkyl of 1 to 6 carbons, alkoxy of 1 to 6carbons, or a pharmaceutically acceptable salt of said compound; withthe proviso: that when R₁₀ has formula (ii) then the claim does notinclude compounds where R₄ is hydrogen and R₁ and R₂ jointly with thenitrogen form a morpholin or a pyrrolidin ring and where R₅ and R₆ bothare H or one of R₅ and R₆ is OCH₃ and the other is H.
 2. A compound inaccordance with claim 1 where R₁₀ represents the formula (i).
 3. Acompound in accordance with claim 1 where R₁₀ represents the formula(ii).
 4. A compound in accordance with claim 2 having the formula

where R₅ and R₆ are independently selected from H, alkyl and alkoxy andR₄ is H or CO—R₈ or a pharmaceutically acceptable salt of said compound.5. A compound in accordance with claim 4 having the formula

or a pharmaceutically acceptable salt of said compound.
 6. A compound inaccordance with claim 3 having the formula

where R₃ is CO—R₇ or CO—O—R₇, R₄ is CO—R₈ and R₅ and R₆ areindependently selected from H, alkyl of 1 to 6 carbons and alkoxy of 1to 6 carbons or any other pharmaceutically acceptable salt of saidcompound.
 7. A compound in accordance with claim 6 having the formula

or any other pharmaceutically acceptable salt of said compound.
 8. Acompound of the formula

or any other pharmaceutically acceptable salt of said compound.
 9. Acompound of the formula

or any other pharmaceutically acceptable salt of said compound.
 10. Amethod of treating a mammal in need of such treatment with apharmaceutical composition having an analgesic effect, the compositionincluding a compound in accordance with claim
 1. 11. A method oftreating a mammal in need of such treatment with a pharmaceuticalcomposition having an analgesic effect, the composition including acompound in accordance with claim
 2. 12. A method of treating a mammalin need of such treatment with a pharmaceutical composition having ananalgesic effect, the composition including a compound in accordancewith claim
 3. 13. A method of treating a mammal in need of suchtreatment with a pharmaceutical composition having an analgesic effect,the composition including a compound in accordance with claim
 4. 14. Amethod of treating a mammal in need of such treatment with apharmaceutical composition having an analgesic effect, the compositionincluding a compound in accordance with claim
 5. 15. A method oftreating a mammal in need of such treatment with a pharmaceuticalcomposition having an analgesic effect, the composition including acompound in accordance with claim
 6. 16. A method of treating a mammalin need of such treatment with a pharmaceutical composition having ananalgesic effect, the composition including a compound in accordancewith claim
 7. 17. A method of treating a mammal in need of suchtreatment with a pharmaceutical composition having an analgesic effect,the composition including a compound in accordance with claim
 8. 18. Amethod of treating a mammal in need of such treatment with apharmaceutical composition having an analgesic effect, the compositionincluding a compound in accordance with claim
 9. 19. A pharmaceuticalcomposition for treating pain in a mammal, the composition comprising acompound in accordance with claim 1 and a pharmaceutically acceptableexcipient.